A Cassette Approach for the Identification of Intein Insertion Sites.
A Cassette Approach for the Identification of Intein Insertion Sites.
复制标题
鉴定内含肽插入位点的盒式方法。
DOI：
10.1007/978-1-4939-6451-2_16
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Sonntag,Tim
中科院分区：
文献类型：
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作者：
Sonntag,Tim
在过去的十年中，分裂蛋白已经成为蛋白质工程、蛋白质半合成和蛋白质功能控制方法的有力工具。它们的主要优势在于蛋白质反式剪接（PTS）反应，该反应使两个独立的，甚至是合成的肽前体能够在翻译后组装蛋白质。然而，由于大多数分裂蛋白的应用都是处理蛋白质的分裂和修饰，因此可能会出现各种问题，从稳定性降低到蛋白质折叠损伤。在本章中，我将讨论如何使用DNA编码的内嵌蛋白磁带来简化和加速鉴定新型感兴趣蛋白（POI）中的功能性分裂内嵌蛋白插入位点。
Over the past decade split inteins have established themselves as powerful tools for protein engineering, protein semisynthesis, and protein functional control approaches. Their key advantage lies in the proteintrans-splicing (PTS) reaction that enables posttranslational protein assembly from two independent, even synthetic, peptide precursors. However, since most split intein applications deal with fragmentation and modification of proteins, various issues can arise, ranging from reduced stability to impairment of protein folding. In this chapter I address how the usage of DNA encoded intein cassettes can streamline and speed up the identification of functional split intein insertion sites in novel proteins of interest (POI).